This invention relates to novel compounds that promote the release of growth hormones when introduced to animals, preferably humans, and methods of use thereof.
The elevation of growth hormone (GH) levels in animals, e.g., mammals including humans, upon administration of GH-releasing compounds can lead to enhanced body weight and to enhanced milk production if sufficiently elevated GH levels occur upon administration. Further, it is known that the elevation of growth hormone levels in mammals and humans can be accomplished by application of known growth hormone releasing agents, such as the naturally occurring growth hormone releasing hormones.
The elevation of growth hormone levels in mammals can also be accomplished by application of growth hormone releasing peptides (GHRPs), some of which have been previously described, for example, in U.S. Pat. Nos. 4,223,019; 4,223,020; 4,223,021; 4,224,316; 4,226,857; 4,228,155; 4,228,156; 4,228,157; 4,228,158; 4,410,512; 4,410,513.
Antibodies to the endogenous growth hormone release inhibitor, somatostatin (SRIF) have also been used to cause elevated GH levels. In this latter example, growth hormone levels are elevated by removing the endogenous GH-release inhibitor (SRIF) before it reaches the pituitary, where it inhibits the release of GH.
These methods for promoting the elevation of growth hormone levels frequently involve materials which are expensive to synthesize and/or difficult to isolate in sufficient purity for administration to a target animal. Low molecular weight, relatively simple and inexpensive compounds that have the ability to promote the release of growth hormone would be desirable in that they could be readily and inexpensively prepared, easily modified chemically and/or physically, as well as easily purified and formulated, and designed to have improved transport properties.
GH and/or GHRPs have been administered to stimulate growth hormone production and/or release, for example, to stimulate growth, enhance milk production, enhance body weight, increase rate of protein synthesis, reduce rate of carbohydrate utilization, increase mobilization of pre-fatty acids. Although the use of many of these compounds such as a series of short peptides (e.g., U.S. Pat. Nos. 5,663,146 and 5,486,505) have been important steps in the design and delivery of compounds having GH and/or GHRP properties, improvements can still be made. For example, improvements can be made in the areas of oral bioavailability, serum retention time, etc.
Non-peptidal or hybrid-peptidal secretagogues have also been described. See U.S. Pat. Nos. 5,494,919; 5,492,920; 5,492,916; 5,622,973; WO95/13069, WO96/15148; WO96/35713; WO97/22367; WO97/00894; WO97/07117; and WO97/11697. Despite the general descriptions of such compounds, it is not possible to make broad generalizations about which particular compounds are favorable. Although some secretagogues, which can promote the release and elevation of growth hormone levels in the blood, have been described, corresponding data on the biological activity has often been lacking. Moreover, even in terms of tripeptides with or without C-terminal modifications, the data suggests that it has heretofore been impossible to make the broad sweeping generalization made in those publications about what would or would not be a favorable amino acid combination at the three positions of a tripeptide holding the C-terminal constant or holding the peptidal portion constant while making changes, or changing the chemical moieties added. Changes in any of the constituents can have great effects on activity. It is submitted that these references do not lead to general teachings of biological efficacy.
In order to maximize the ability to select and tailor a compound, it would be desirable to have a class of compounds that generally provide good growth hormone releasing effects and have at least one other desirable biological activity such as better bioavailability, absorption, metabolism, pharmacokinetics, excretions, etc. It would also be desirable to have compounds which can promote the release and elevation of growth hormone levels in the blood of animals, particularly in humans, to be able to use such compounds to promote the release and/or elevation of growth hormone levels in the blood of animals and humans, and to provide methods for promoting the release and/or elevation of growth hormone levels in the blood of animals using such compounds.
The aforementioned discussion illustrates that a broad chemical diversity of synthetic GHRPs ranging from peptides to partial peptides to non-peptides. Overall, the peptides and partial peptides have been the most effective in promoting elevated growth hormone levels. For example, partial peptides consisting of natural and unnatural amino acids of different chain lengths and C-terminal amide groups or a substituted amide with various organic chemical groups. Results published as early as 1982 stated that certain GHRPs with only 3-7 amino acids released GH and that having a D-amino acid at certain positions was useful. From 1982 to the present, GHRPs with more potent GH releasing activity have been developed. This research taught that certain amino acid positions could have certain substitutions but not others, and that one amino acid residue could affect what other substitutions could be made.
Until compounds having the optimum physical-chemical properties and physiological-biological actions and effects are discovered for various diagnostic and therapeutic uses in humans, it is important to discover a general chemical approach that will result in new types of GHRPs. Such a broader GHRP chemical base will make it possible to better implement and refine the GHRP approach.
Properties of GHRPs that are important include that they are effective when administered orally. In addition, the compound should augment the normal pulsatile physiological secretion of GH. In some subjects with decreased GH secretion, GH can be replaced in a physiological way. Physiological replacement of a hormonal deficiency improves health while minimizing the potential adverse action of the hormone. This is especially important in treating older men and women, as they may be particularly susceptible to the adverse effects of over-treatment with GH. Already, chronic administration of GHRPs to animals and humans has produced anabolic effects. Body weight gain has been increased in rats, milk production has been increased in cows. Additionally, when a compound such as DAla-Dxcex2Nal-Ala-Trp-DPhe-Lys-NH2 (GHRP-2) was administered to short-statured children with various degrees of GH deficiency 2-3 times per day over a 2 year period, the rate of height velocity has been accelerated in those children.
In principle, the anabolic biological effects of GHRPs emphasize the potential clinical value of the GHRP approach. The finding that GHRP-2 is less effective on height velocity than usually obtained with chronic recombinant human growth hormone (rhGH) administration, underscores the desirability for improving the GHRP approach. This includes further optimization and extension of the range of the GHRP chemistry in order to produce more effective biological actions.
In looking at these compounds, one looks at a varied series of biological effects such as the duration of action of GHRP. Other parameters that may substantially be affected by the chemistry of the GHRP include desensitization of the GHRP GH response, actions on the hypothalamus, effects on SRIF release and action, effects on ACTH and PRL release as well as possible effects on putative subclasses of GHRP receptors. All of these actions are directly and/or indirectly dependent on the GHRP chemistry, pattern and efficiency of oral absorption as well as the metabolism and secretion of the particular GHRP.
We have now discovered a new group of compounds (sometimes referred to as secretagogues) that provide desirable in vitro and in vivo growth hormone releasing activity and have at least one other desirable biological activity such as increased retention time. These compounds have the following formulas:
A1-A2-Xxe2x80x83xe2x80x83Formula I
wherein
A1 is Aib (aminoisobutyric acid), inip (isonipecotyl) or ABU (aminobutyric acid). The Aib residue can be substituted or unsubstituted. Preferred substituents include C1-C6 alkyl and halogens. Aib is preferably unsubstituted. Aib is preferably xcex1Aib. ABU is preferably xcex3ABU or xcex1xcex3ABU, more preferably xcex1,xcex3ABU;
A2 is any natural L-amino acid or Pal, or their respective D-isomers, Dxcex1Nal (xcex1-naphthyl-D-alanine) or Dxcex2Nal (xcex2-naphthyl-D-alanine), preferably A2 is DTrp, Dxcex1Nal (xcex1-naphthyl-D-alanine) or Dxcex2Nal (xcex2-naphthtyl-D-alanine), more preferably
A2 is DTrp or Dxcex1Nal;
X is
(1) R1-R2-Z, wherein R1 and R2 are any natural L-amino acid, Pal, xcex1Nal, xcex2Nal, DpCl, CHx, where CHx is cyclohexyl, CHxAla, or any of their respective D-isomers, preferably R1 is DPro, DTrp, Dxcex2Nal or DPhe, more preferably R1 is DPro or DTrp; and R2 is preferably Gly, Phe, Pro, DPro, DPhe, DPal, DLeu, DHis, DVal, DGln, DArg, DAla, DSer, DThr, DIle, Arg, Orn Lys, Ala, Pal, Thr, Val, PheCHx, CHxAla or CHx, where x is preferably 1-8, more preferably 1 to 5; and Z is CONH2 or COOH;
(2) DpR3Phe-R4-Z, wherein R3 is a halogen, preferably Cl, and R4 is any natural L-amino acid or Pal, or their respective D-isomers, preferably R4 is Phe or Arg, and Z is CONH2 or COOH;
(3) NH(CH2)nNH, where n is 1 to 8, such as -2-aminoethylamide, -3-aminopropylamide, -4-aminobutylamide, -5-aminopentylamide, or -6-aminohexylamide;
(4) R5-R6, wherein R5 is any natural L-amino acid, Pal, xcex1Nal, xcex2Nal, DpCl, CHx where x is 1 to 10, or any of their respective D-isomers, preferably R5 is DPro or DTrp, and R6 is
(a) diisobutylamide
(b) dipropylamide
(c) butylamide
(d) pentylamide
(e) dipentylamide
(f) C(xe2x95x90O) (substituted heteroalicyclic or heteroaromatic) such as
-piperidine-3-methylbenzylether
-N-diethylnipectamide
-N-piperazine methylsulfonamide
-diethylamide
-m-methylpiperidine
-3,3-diphenylpropylamide
-4-piperidino piperidinamide
-4-phenyl-piperidinamide
-N-methylpiperazine
-2-morpholinoethylamine
-spiroindole methylsulfonamide
-pyrrolidine amide
-indoleamide
-3-piperidine methanolamide
-tropin amide
-2-aminoethylamide
-3-aminopropylamide
-4-aminobutylamide
-5-aminopentylamide
-6-aminohexylamide;
(5) DTrp Phe ArgR7, wherein R7 is NH(CH2)nNH, where n is 1 to 8, such as -2-aminoethylamide, -3-aminopropylamide, -4-aminobutylamide, -5-aminopentylamide, or -6-aminohexylamide; or
(6) R8-R9-R10-Z, wherein R8 is DTrp, DPro, Dxcex1Nal or Dxcex2Nal, preferably R8 is DTrp or DPro, R9 is any natural L-amino acid or Pal, or their respective D-isomers, preferably R9 is Phe, DVal, DPro, DIle, Ile, more preferably Ris Phe, DVal or DPro; R10 is any natural L-amino acid or Pal, or their respective D-isomers, preferably R10 is Lys or Arg, and Z is CONH2 or COOH, preferably Z is CONH2.
A1xe2x80x2-Xxe2x80x2xe2x80x83xe2x80x83Formula II:
xe2x80x83wherein A1xe2x80x2 is Aib, inip, ABU, IMC (imidazole carboxylic acid), Ava, 4-IMA (Nxcex1-imidazole acetic acid), xcex2Ala, Ileu, Trp, His, DpCl, CHx, or any of their respective D-isomers. The Aib residue can be substituted or unsubstituted. Preferred substituents include N- and N-,N-C1-C6 alkyl, halogens, N- and N-,N-2 hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl and 3-hydroxyisobutyl. Aib is preferably unsubstituted. Aib is preferably xcex1Aib. ABU is preferably xcex3ABU or xcex1xcex3ABU, more preferably xcex1,xcex3ABU; and
Xxe2x80x2 is
(1) R1xe2x80x2-R2xe2x80x2-Z, wherein R1xe2x80x2 is any natural L-amino acid or Pal, or their respective D-isomers, Dxcex1Nal or Dxcex2Nal, preferably R1xe2x80x2 is DTrp, Dxcex1Nal or Dxcex2Nal, more preferably R1xe2x80x2 is DTrp or Dxcex1Nal, and R2xe2x80x2 is any natural L-amino acid, Pal, xcex1Nal, xcex2Nal, DpCl, Aib, preferably xcex1Aib, CHx where x is 1 to 10, or CHxAla, or any of their respective D-isomers, and Z is CONH2 or COOH, preferably Z is CONH2; or
(2) R3xe2x80x2-R4xe2x80x2, wherein R3xe2x80x2 is any natural L-amino acid or Pal, or their respective D-isomers, Dxcex1Nal or Dxcex2Nal, preferably R3 is DPro, DTrp, Dxcex1Nal or Dxcex2Nal, more preferably R3xe2x80x2 is DPro, DTrp or Dxcex1Nal, and R4xe2x80x2 is NH(CH2)nNH, where n is 1 to 8, such as -2-aminoethylamide, -3-aminopropylamide, -4-aminobutylamide, -5-aminopentylamide, or -6-aminohexylamide.
The organic and inorganic addition salts thereof are also included.
In an alternative embodiment the compound has the formula
A1xe2x80x3-Y,xe2x80x83xe2x80x83Formula III:
wherein
A1xe2x80x3 is Aib, inip, ABU, xcex2Ala, His, Sar or any of their respective D-isomers. The Aib residue can be substituted or unsubstituted. Preferred substituents include N- and N-,N-C1-C6 alkyl, halogens, N- and N-,N-2 hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl and 3-hydroxyisobutyl. Aib is preferably unsubstituted. A1xe2x80x2, is preferably Aib, inip or ABU. More preferably Aib is xcex1Aib. Abu is preferably xcex3Abu or xcex1,xcex3Abu, more preferably xcex1,xcex3Abu.
Y is A2xe2x80x2-A3-A4-A5-A6-Zxe2x80x2,
A2xe2x80x2-A3-A4-A5-Zxe2x80x2 or A2xe2x80x2-A3-A4-Zxe2x80x2
wherein A2xe2x80x2 is A5-A2xe2x80x3 or A2xe2x80x3,
wherein A5 is a spacer amino acid such as His,
A2xe2x80x3 is as defined above for A2. A2xe2x80x3 is preferably DTrp, Dxcex1Nal or Dxcex2Nal. A2xe2x80x3 is more preferably DTrp.
A3, A4 and A5 are any natural L-amino acid, Pal, xcex1Nal, xcex2Nal, Nle, Arg-DPro, DPCl, D or L (CHX), cyclohexylalanine (CHXAla), or any of their respective D-isomers, preferably A3 is DPro, DTrp, Dxcex2Nal or DPhe, more preferably A3 is DPro or DTrp; and A4 is preferably Gly, Phe, Pro, Ile, DPro, DPhe, DPal, DLeu, DHis, DVal, DGln, DIle, DNle, DArg, DAla, DSer, DThr, DIle, Arg, Orn Lys, Ala, Pal, Thr, Val, PheCHX, CHXAla or CHX. A4 is preferably DSer, DAug, DPro, DTrp, DVal, DIle, DThr, DNVal, DNle, Ile, Pro, Phe and still more preferably, A4 is DPro. A5 is preferably Ile, Arg, Pal, DArg, DSer, Lys and Arg-DPro. More preferably A5 is Arg, DArg, and Lys.
Zxe2x80x2 is NH2, OH or alkylamino or aminoalkylamino, preferably the alkylamino is NH (C1-C10 alkyl) e.g. NH(CH2)nCH3, where n is 1 to 10 such as 
xe2x80x83N di-(C1-C10 alkyl) e.g., N di-(CH2)nCH3 such as 
xe2x80x83preferably the aminoalkylamino is a NH (C1-C10 alkylamino, e.g. NH(CH2)nNH2 such as 
N (di C1-C10 alkylamino), e.g., N [di-(CH2)nNH2] such as 
These compounds can be administered to an animal to promote release of serum growth hormone levels. Thus, these secretagogues can be used in a range of methods for example, to increase milk production, enhance body growth, treat hypothalmic pituitary dwarfism, osteoporosis, burns and renal failure, and to promote wound healing. They can also be used diagnostically. For example, to discover a loss of growth hormone receptor functioning.
The compounds described herein are typically easy to synthesize, have efficacy at promoting an increase in serum growth hormone levels, and are desirable for large scale production and utilization. In addition, these compounds may be advantageous in having physiochemical properties which are desirable for the efficient delivery of such polypeptide compounds to a wide variety of animal species because of an improvement in at least one of bioavailability, absorption, metabolism, pharmacokinetics and excretion. The preferred methods of delivery are oral, nasal and continuous delivery utilizing special chemical/mechanical methods of delivery. Pulsed therapy is one preferred method of administration. These compounds have either of the following two formulas:
A1-A2-Xxe2x80x83xe2x80x83Formula I:
wherein
A1 is Aib (aminoisobutyric acid), inip (isonipecotyl) or ABU (aminobutyric acid). The Aib residue can be substituted or unsubstituted. Preferred substituents include C1-C6 alkyl and halogens. Aib is preferably unsubstituted. Aib is preferably xcex1Aib. ABU is preferably xcex3ABU or xcex1xcex3ABU, more preferably xcex1,xcex3ABU;
A2 is any natural L-amino acid or Pal, or their respective D-isomers, Dxcex1Nal (xcex1-naphthyl-D-alanine) or Dxcex2Nal (xcex2-naphthtyl-D-alanine), preferably A2 is DTrp, Dxcex1Nal (xcex1-naphthyl-D-alanine) or Dxcex2Nal (xcex2-naphthyl-D-alanine), more preferably A2 is DTrp or Dxcex1Nal;
X is
(1) R1-R2-Z, wherein R1 and R2 are any natural L-amino acid, Pal, xcex1Nal, xcex2Nal, DpCl, CHx, CHxAla, or any of their respective D-isomers, preferably R1 is DPro, DTrp, Dxcex2Nal or DPhe, more preferably R1 is DPro or DTrp; and R2 is preferably Gly, Phe, Pro, DPro, DPhe, DPal, DLeu, DHis, DVal, DGln, DArg, DAla, DSer, DThr, DIle, Arg, Orn Lys, Ala, Pal, Thr, Val, PheCHx, CHxAla or CHx, where x is preferably 1-8, more preferably 1 to 5; and Z is CONH2 or COOH;
(2) DpR3Phe-R4-Z, wherein R3 is a halogen, preferably Cl, and R4 is any natural L-amino acid or Pal, or their respective D-isomers, preferably R4 is Phe or Arg, and Z is CONH2 or COOH;
(3) NH(CH2)nNH, where n is 1 to 8, such as -2-aminoethylamide, -3-aminopropylamide, -4-aminobutylamide, -5-aminopentylamide, or -6-aminohexylamide;
(4) R5-R6, wherein R5 is any natural L-amino acid, Pal, xcex1Nal, xcex2Nal, DpCl, CHx where x is 1 to 10, or any of their respective D-isomers, preferably R5 is DPro or DTrp, and R6 is
(a) diisobutylamide
(b) dipropylamide
(c) butylamide
(d) pentylamide
(e) dipentylamide
(f) C(xe2x95x90O)(substituted heteroalicyclic or heteroaromatic) such as
-piperidine-3-methylbenzylether
-N-diethylnipectamide
-N-piperazine methylsulfonamide
-diethylamide
-m-methylpiperidine
-3,3-diphenylpropylamide
-4-piperidino piperidinamide
-4-phenyl-piperidinamide
-N-methylpiperazine
-2-morpholinoethylamine
-spiroindole methylsulfonamide
-pyrrolidine amide
-indoleamide
-3-piperidine methanolamide
-tropin amide
-2-aminoethylamide
-3-aminopropylamide
-4-aminobutylamide
-5-aminopentylamide
-6-aminohexylamide;
(5) DTrp Phe Arg R7, wherein R7 is NH(CH2)nNH, where n is 1 to 8, such as -2-aminoethylamide, -3-aminopropylamide, -4-aminobutylamide, -5-aminopentylamide, or -6-aminohexylamide; or
(6) R8-R9-R10-Z, wherein R8 is DTrp, DPro, Dxcex1Nal or Dxcex2Nal, preferably R8 is DTrp or DPro, R9 is any natural L-amino acid or Pal, or their respective D-isomers, preferably R9 is Phe, DVal, DPro, DIle, Ile, more preferably R9 is Phe, DVal or DPro; R10 is any natural L-amino acid or Pal, or their respective D-isomers, preferably R10 is Lys or Arg, and Z is CONH2 or COOH, preferably Z is CONH2.
A1xe2x80x2-Xxe2x80x2xe2x80x83xe2x80x83Formula II:
xe2x80x83wherein A1xe2x80x2, is Aib, inip, ABU, IMC (imidazole carboxylic acid), Ava, 4-IMA (Nxcex1-imidazole acetic acid), xcex2Ala, Ileu, Trp, His, DpCl, CHx, or any of their respective D-isomers. The Aib residue can be substituted or unsubstituted. Preferred substituents include N- and N-,N- C1-C6 alkyl, halogens, N- and N-,N-2 hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl and 3-hydroxyisobutyl. Aib is preferably unsubstituted. Aib is preferably xcex1Aib. ABU is preferably xcex3ABU or xcex1xcex3ABU, more preferably xcex1,xcex3ABU; and
Xxe2x80x2 is
(1) R1xe2x80x2-R2xe2x80x2-Z, wherein R1xe2x80x2 is any natural L-amino acid or Pal, or their respective D-isomers, Dxcex1Nal or Dxcex2Nal, preferably R1xe2x80x2 is DTrp, Dxcex1Nal or Dxcex2Nal, more preferably R1 is DTrp or Dxcex1Nal, and R2xe2x80x2 is any natural L-amino acid, Pal, xcex1Nal, xcex2Nal, DpCl, Aib, preferably xcex1Aib, CHx where x is 1 to 10, or CHxAla, or any of their respective D-isomers, and Z is CONH2 or COOH, preferably Z is CONH2; or
(2) R3xe2x80x2-R4xe2x80x2, wherein R3xe2x80x2 is any natural L-amino acid or Pal, or their respective D-isomers, Dxcex1Nal or Dxcex2Nal, preferably R3xe2x80x2 is DPro, DTrp, Dxcex1Nal or Dxcex2Nal, more preferably R3xe2x80x2 is DPro, DTrp or Dxcex1Nal, and R4xe2x80x2 is NH(CH2)nNH, where n is 1 to 8, such as -2-aminoethylamide, -3-aminopropylamide, -4-aminobutylamide, -5-aminopentylamide, or -6-aminohexylamide.
The organic and inorganic addition salts thereof are also included.
The abbreviations for the residues of amino acids used herein are in agreement with the standard nomenclature, and are set forth below:
Gly Glycine
Tyr L-Tyrosine
Ile L-Isoleucine
Glu L-Glutamic Acid
Thr L-Threonine
Moreover, all of the three letter-abbreviations of the amino acids preceded by a xe2x80x9cDxe2x80x9d indicate the dextro-isomer of the aminoacidic residue, and glycine is considered to be included in the term naturally occurring L-amino acids. Other abbreviations used herein include the following:
In one embodiment of the present invention, a group of preferred compounds includes:
xcex3ABUDTrpDTrpArgCOOH
xcex1,xcex3ABUDTrpDTrpArgNH2 
xcex1,xcex3ABUDTrpDTrpOrnNH2 
xcex1,xcex3ABUDxcex1NalDTrpLysNH2 
xcex1,xcex3ABUDxcex1NalDTrpArgNH2 
xcex1,xcex3AbuDxcex1NalDTrpArgNH2 
xcex1AibDTrpDTrpArgNH2 
xcex1AibDxcex1NalDTrpArgNH2 
xcex1AibDTrpDTrpArgCOOH
xcex1AibDxcex1NalDTrpArgCOOH
xcex1AibDxcex1TrpDTrpArgNH2 
xcex1AibDTrpDPheArgNH2 
inipDxcex1NalDTrpPheNH2 
inipDxcex1NalDTrpCHxAlaNH2 
inipDxcex1NalDTrpPheCOOH
inipDxcex1NalDTrpPalNH2 
inipDxcex1NalDTrpThrNH2 
inipDxcex1NalDTrpValNH2 
inipDxcex1NalDxcex2NalPheNH2 
inipDxcex1NalDTrpPheCOOH
inipDxcex2NalDTrpPheNH2 
xcex1 AibDTrpDProGlyNH2 
xcex1 AibDTrpDProPheNH2 
xcex1 AibDTrpDProProNH2 
xcex1AibDTrpDProDProNH2 
xcex1AibDTrpDProDPheNH2 
xcex1AibDTrpDProDPalNH2 
xcex1AibDTrpDProDTrpNH2 
xcex1AibDTrpDProDLeuNH2 
xcex1AibDTrpDProDHisNH2 
xcex1AibDTrpDProDValNH2 
xcex1AibDTrpDProGlnNH2 
xcex1AibDTrpDProArgNH2 
xcex1AibDTrpDProLysNH2 
xcex1AibDTrpDProDAlaNH2 
inipDxcex1NalDpClPhePheNH2 
inipDxcex1NalDpClPheArgNH2 
inipDxcex1NalDTrpDProNH2 
xcex1AibDTrpDProDSerNH2 
xcex1AibDTrpDProDThrNH2 and
xcex1AibDTrpDProDIleNH2.
In another embodiment of the present invention, a group of preferred compounds includes:
inipDTrpDTrpPheLysNH2 
inipDxcex2NalDTrpPheLysNH2 
xcex3ABUDxcex2NalDTrpPheLysNH2 
xcex1,xcex3ABUDTrpDTrpPheLysNH2 
xcex2AlaDTrpDTrpPheLysNH2 
xcex1,xcex3ABUDxcex2NalDTrpPheLysNH2 
xcex1,xcex3ABUDTrpDTrpPheArgNH2 
xcex1,xcex3ABUDxcex1NalDTrpPheArgNH2 
inipDxcex2BNalDTrpPheLysNH2 
inipDTrpDTrpPheArgNH2 
xcex2AlaDxcex1NalDTrpPheArgNH2 
xcex1AibDTrpDTrpPheArgNH2 
xcex1AibDTrpDTrpPheArgCOOH
inipDTrpDTrpPheArgCOOH
inipDxcex1NalDTrpPheArgNH2 
inipDxcex1NalDTrpPheArgCOOH
inipDxcex1NalDxcex2NalPheArgNH2 
inipDxcex1NalDTrpPheDSerNH2 
inipDxcex1NalDTrpPheDThrNH2 
inipDxcex1NalDTrpPheGlyNH2 
inipDxcex1NalDTrpPheGlnNH2 
inipDxcex1NalDTrpPheDGlnNH2 
xcex1AibDxcex1NalDTrpPheGlnNH2 
inipDxcex1NalDTrpPheDHisNH2 
xcex1AibDTrpDProPheArgNH2 
xcex1AibDTrpDProPheDArgNH2 
xcex1AibDTrpDProDValArgNH2 
xcex1AibDTrpDProDValDLysNH2 
xcex1AibDTrpDProDValDArgNH2 
xcex1AibDTrpDProDProArgNH2 
xcex1AibDTrpDProDProDPalNH2 
xcex1AibDTrpDProDProDArgNH2 
xcex1AibDTrpDProDIleDArgNH2 
xcex1AibDTrpDProDIleArgNH2 
xcex1AibDTrpDProDProDLysNH2 and
xcex1AibDTrpDProIleArgNH2.
In the above Formula I, where X is R5-R6 and R6 is a C(xe2x95x90O) (substituted heteroalicyclic or heteroaromatic), the heteroatom is selected from the group consisting of O, N, S and P.
The heteroalicyclic moiety preferably contains 2 to 12 carbon atoms, more preferably 3 to 8 carbon atoms. The heteroaromatic moiety preferably contains 5 to 12 carbon atoms, more preferably 5 to 11 carbon atoms. Substituents include NH2, C1-C12 lower alkyl, and as listed below.
Examples include piperidine-3-methyl-benzylether, N-diethylnipectamide, N-piperazine methylsulfonamide, diethylamide, m-methylpiperidine, 3,3-diphenylpropylamide, 4-piperidino piperidinamide, 4-phenyl-piperidinamide, N-methyl 1-piperiazine, 2-morpholinoethylamine, spiroindole methylsulfonamide, pyrrolidine amide, indoleamide, 3-piperidine methanol amide, tropin amide, 2-aminoethylamide, 3-aminopropylamide, 4-aminobutylamide, 5-aminopentylamide, 6-aminohexylamide. Preferred substituted heteralicyclic or heteroaromatic include N-diethylnipectamide, piperidine-3-methyl-benzylether, N-piperazine methyl sulfonamide, diethylamide and m-methylpiperidine. Even more preferred are N-diethylnipectamide and piperidine-3-methyl-benzylether.
Preferably, the compound has the structure AibDTrpX, where X is DProNH2, DPro-diisobutylamide, DProbutylamide, DPro-C(xe2x95x90O)(substituted heteroalicyclic or heteroaromatic), and DTrp-Phe-Arg-5-aminopentamide and organic and inorganic addition salts thereof. More preferably, X is DPro-diisobutylamide, DPro-C(xe2x95x90O)(substituted heteroalicyclic or heteroaromatic) and DTrp PheArg-5-aminopentamide, and organic and inorganic addition salts thereof. Still more preferably, X is DPro-diisobutylamide or DTrp-Phe-Arg-5-aminopentamide, and organic and inorganic addition salts thereof. Even more preferably, X is DPro-diisobutylamide and organic and inorganic addition salts thereof.
In an alternative embodiment the compound has the formula
A1xe2x80x3-Y,
wherein
A1xe2x80x3 is Aib, inip, ABU, xcex2Ala, His, Sar or any of their respective D-isomers. The Aib residue can be substituted or unsubstituted. Preferred substituents include N- and N-,N-C1-C6 alkyl, halogens, N- and N-,N-2 hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl and 3-hydroxyisobutyl. Aib is preferably unsubstituted. A1xe2x80x3 is preferably Aib, inip or ABU. More preferably Aib is xcex1Aib. Abu is preferably xcex3Abu or xcex1,xcex3Abu, more preferably xcex1,xcex3Abu.
Y is A2xe2x80x2-A3-A4-A5-A6-Zxe2x80x2,
A2xe2x80x2-A3-A4-A5-Zxe2x80x2 or A2xe2x80x2-A3-A4-Zxe2x80x2
wherein A2xe2x80x2 is A5-A2xe2x80x3 or A2xe2x80x3,
wherein A5 is a spacer amino acid such as His,
A2xe2x80x3 is as defined above for A2. A2xe2x80x3 is preferably DTrp, Dxcex1Nal or Dxcex2Nal. A2xe2x80x3 is more preferably DTrp.
A3, A4 and A5 are any natural L-amino acid, Pal, xcex1Nal, xcex2Nal, Nle, Arg-DPro, DPCl, D or L (CHX), cyclohexylalanine (CHXAla), or any of their respective D-isomers, preferably A3 is DPro, DTrp, Dxcex2Nal or DPhe, more preferably A3 is DPro or DTrp; and A4 is preferably Gly, Phe, Pro, Ile, DPro, DPhe, DPal, DLeu, DHis, DVal, DGln, DIle, DNle, DArg, DAla, DSer, DThr, DIle, Arg, Orn Lys, Ala, Pal, Thr, Val, PheCHX, CHXAla or CHX. A4 is preferably DSer, DAug, DPro, DTrp, DVal, DIle, DThr, DNVal, DNle, Ile, Pro, Phe and still more preferably, A4 is DPro. A5 is preferably Ile, Arg, Pal, DArg, DSer, Lys and Arg-DPro. More preferably A5 is Arg, DArg, and Lys.
Zxe2x80x2 is NH2, OH or (aminoalkyl) or (aminoalkylamino), preferably the aminoalkyl is NH (C1-C10 alkyl) e.g. NH(CH2)nCH3, where n is 1 to 10 such as 
xe2x80x83N di-(C1-C10 alkyl) e.g., N di-(CH2)n CH3 such as 
xe2x80x83preferably the alkylamino is a NH (C1-C10 alkylamino, e.g. NH(CH2)nNH2 such as 
xe2x80x83N (di C1-C10 alkylamino), e.g., N [di-(CH2)nNH2] such as 
Preferred examples include moieties such as -2-aminoethylamide, -3-aminopropylamide, -4-aminobutylamide, -5-aminopentylamide, or -6-aminohexylamide; N-dimethylamide; N-diethylamide; N-dipropylamide; N-dibutylamide; N-diisobutylamide; N-dipentylamide; N-dihexylamide;
A particularly preferred embodiment is Aib-Y, more preferably xcex1Aib-Y.
Y is preferably A2xe2x80x3-DPro-A4-A5-A6-Zxe2x80x2; A2xe2x80x3-A3-A4-Zxe2x80x2; or A2xe2x80x3-A3-A4-A5Zxe2x80x2. Y is more preferably A2xe2x80x3-DPro-A4-Zxe2x80x2 or A2xe2x80x3-DPro-A4-Zxe2x80x2 or A2xe2x80x3-DPro-A4-A5-Zxe2x80x2. Still more preferably Y is A2xe2x80x3-DPro-A4-A5-Zxe2x80x2. Zxe2x80x2 is preferably -NH2.
Preferred embodiments include
xcex1Aib-DTrp-DPro-A4-A5-A6-Zxe2x80x2;
xcex1Aib-DTrp-DPro-A4-A5-Zxe2x80x2;
xcex1Aib-DTrp-DPro-A4-Zxe2x80x2;
xcex1Aib-DTrp-DPro-A4-Arg-NH2;
xcex1Aib-DTrp-DPro-A4-Arg-A6-NH2;
xcex1Aib-DTrp-DPro-A4-Arg-Gly-NH2;
xcex1Aib-Dxcex1Nal-DPro-A4-A5-A6-Zxe2x80x2;
xcex1Aib-Dxcex1Nal-DPro-A4-A5-Zxe2x80x2;
xcex1Aib-Dxcex1Nal-DPro-A4-Zxe2x80x2;xe2x80x2
xcex1Aib-Dxcex1Nal-DPro-A4-NH2;
xcex1Aib-Dxcex1Nal-DPro-A4-Arg-NH2;
and xcex1Aib-Dxcex1Nal-DPro-A4-Arg-Gly-NH2.
A4 is preferably DIle, DThr, DNle, DVal, DGln, DAla, DPhe, DTrp, DNVal and Arg.
Exemplery representatives of xcex1Aib-A2xe2x80x3-DPro-A4-Arg-Zxe2x80x2 include
xcex1AibDTrpDProDIleArgNH2;
xcex1AibDTrpDProDThrArgNH2;
xcex1AibDTrpDProDValArgNH2;
xcex1AibDTrpDProDNleArgNH2; and
xcex1AibDxcex1NalDProDIleDArgNH2.
Exemplary representatives of:
xcex1Aib-A2xe2x80x3-DPro-A4-Z include
xcex1Aib-DTrp-DPro-DThr-NH2;
xcex1Aib-DTrp-DPro-DGln-NH2;
xcex1Aib-DTrp-DPro-Arg-NH2;
xcex1Aib-DTrp-DPro-DAla-NH2;
xcex1Aib-DTrp-DPro-DPhe-NH2;
xcex1Aib-DTrp-DPro-DTrp-NH2;
xcex1Aib-DTrp-DPro-DVal-NH2;
xcex1Aib-DTrp-DPro-DNVal-NH2; and
xcex1Aib-DTrp-DPro-DIle-NH2;
Exemplary representatives of xcex1Aib-A2xe2x80x3-DPro-A4-Arg-A6-Z include compounds of the formula xcex1Aib-A2xe2x80x3-DPro-A4-Arg-Gly-NH2 such as
xcex1Aib-DTrp-DPro-DIle-Arg-Gly-NH2;
xcex1Aib-DTrp-DPro-DThr-Arg-Gly-NH2; and
xcex1Aib-DTrp-DPro-DNle-Arg-Gly-NH2.
Representative compounds are set forth below:
inipDxcex1NalDTrpNH2;
inipDxcex1NalDValNH2;
xcex1AibDTrpDValNH2;
xcex1AibDTrpDProDSerNH2;
xcex1AibDTrpDProDArgNH2;
xcex1AibDTrpDProDPheNH2;
xcex1AibDTrpDProDTrpNH2;
xcex1AibDTrpDValDValNH2;
xcex1AibDValDProDValNH2;
xcex1AibDValDValDValNH2;
xcex1AibDTrpDProDLysNH2;
xcex1AibDProDProDValNH2;
inipDxcex1NalDTrpDValNH2;
xcex1AibDTrpDProIleNH2;
xcex1xcex3AbuDxcex1NalDTrpDIleNH2;
inipDxcex1NalDTrpDProIleNH2;
inipDxcex1NalDTrpPheIleNH2;
inipDxcex1NalDTrpDValArgNH2;
xcex1AibDTrpDProDValDValNH2;
xcex1AibDTrpDProDProDPalNH2;
xcex1AibDTrpDProDValArgDProNH2;
xcex1AibDTrpDProDIleDArgNH2;
xcex1xcex3AbuDTrpDTrpDIleNH2;
inipDxcex1NalDTrpPheDValNH2;
xcex1AibDTrpDProValNH2;
xcex1AibDTrpDIleDIleNH2;
xcex1AibDTrpDProLeuNH2;
xcex1AibDTrpDProThrNH2;
DHisDTrpDProDValArgNH2;
DHisDTrpDProDThrNH2;
xcex1AibDTrpDProDIleNH2;
xcex1AibDTrpDPheDValNH2;
xcex1AibDTrpDProDValDArgNH2;
xcex1AibDTrpDProDAlaNH2;
xcex1AibDTrpDProDProNH2;
xcex1AibDTrpDProArgNH2;
xcex1AibDTrpDProDValNH2;
inipDxcex1NalDTrpDProNH2;
xcex1AibDxcex1NalDProDValDArgNH2;
xcex1AlbDxcex1NalDProDIleDArgNH2;
xcex1AibDTrpDProDProDLysNH2;
xcex1AibHisDxcex1NalDPheLysNH2;
xcex1AibHisDTrpDProDValNH2;
xcex1AibHisDTrpDProDIleNH2;
xcex1AibHisDTrpDProValArgNH2;
xcex1AibHisDTrpDProDValArgNH2;
xcex1AibDxcex1NalDProDValNH2;
xcex1AibDTrpDProDThrArgNH2;
xcex1AibDTrpDProDNleArgNH2;
xcex1AibDTrpDProDNValArgNH2;
xcex1AibDTrpDProIleArgNH2;
xcex1AibDTrpDProDProArgNH2;
xcex1AibDTrpDProProArgNH2;
xcex1AibDTrpDProDProDArgNH2;
xcex1AibDTrpDProDIleArgNH2;
xcex1AibDTrpDProPheDSerNH2;
xcex1AibDTrpDProPheArgNH2;
xcex1AibDTrpDProDValArgNH2;
SarDTrpDTrpPheArgNH2;
xcex1AibDxcex1NalDProDProArgNH2;
xcex1AibDxcex1NalDProDNValArgNH2;
xcex1AibDxcex1NalDProDIleArgNH2;
xcex1AibDxcex1NalDProDValLysNH2;
xcex1AibDxcex1NalDProDThrArgNH2;
xcex1AibDxcex1NalDProDThrArgNH2;
xcex1AibDxcex1NalDProDValArgNH2;
xcex1AibDxcex1NalDProDValArgNH2;
xcex1AibDTrpDProDNleNH2;
xcex1AibDTrpDProDNValNH2.
xcex1AibDTrpDProDIle-Xa, where Xa is
2-aminoethylamnide,
5-aminopentylamide, or
3-aminopropylamide.
xcex1AibDTrpDProDVal-Xb, where Xb is
2-aminoethylamide,
dimethylamide, or
diethylamide.
xcex1AibDTrpDProDPro-Xc, where Xc is
2-aminoethylamide.
The following compounds are preferred
xcex1AibDTrpDProDIleXd, where Xd is
5-aminopentylamide,
3-aminopropylamide,
2-aminoethylamide, or
4-aminobutylamide
xcex1AibDTrpDProDValXe, where Xe is
N-dimethylamide,
N-diethylamide, or
2-aminoethylamide;
xcex1AibDTrpDProDValXf, where Xf is
5-aminopentylamide;
xcex1AibDTrpDProDNleXg, where Xg is
5-aminopentylamide;
xcex1AibDTrpDProDProArgNH2;
xcex1AibDTrpDProDValDArgNH2;
xcex1AibDTrpDProDValArgNH2;
xcex1AibDTrpDProDIleArgNH2;
xcex1AibDxcex1NalDProDValArgNH2;
xcex1AibDxcex1NalDProDValArgNH2;
xcex1AibDxcex1NalDProDIleArgNH2;
xcex1AibDxcex1NalDProDValLysNH2;
inipDxcex1NalDxcex1NalPheArgNH2;
xcex1AibDTrpDProDThrArgNH2;
xcex1AibDTrDProDNleArgNH2;
xcex1AibDTrpDProDNValArgNH2;
xcex1AibDTrpDProDIleArgGlyNH2;
xcex1AibDTrpDProDProDIleArgGlyNH2;
xcex1AibDTprDProDNleArgGlyNH2; and
xcex1AibDTrpDProDThrArgGlyNH2;
In one embodiment one uses compound from compounds having the formula
xcex1AibDTrpDProDProA4ArgNH2 or
xcex1AibDTrpDProDProA4ArgGlyNH2.
Preferred examples are selected from the group consisting of
xcex1AibDTrpDProDIleArgNH2 
xcex1AibDTrpDProDIleArgGlyNH2 
xcex1AibDTrpDProDProDIleArgNH2, and
xcex1AibDTrpDProDProDIleArgGlyNH2.
In an alternate embodiment, the following peptides are of interest:
Dxcex2NalAlaTrpDPheLysGlnGlyNH2 
DAlaDTrpAlaTrpDPheLysValGlyNH2 
DAlaDxcex2NalAlaTrpDPheLysGlnGlyGlyGlyNH2 
DAlaDTrpAlaTrpDPheLysHisGlyNH2
These secretagogues can be used therapeutically for any use for which growth hormone can be used, such as treating hypothalamic pituitary dwarfism, osteoporosis, burns, and renal failure for acute use, for non-union bone fracture, and to promote wound healing. Additionally, it can be used to promote recovery from surgery, and acute/chronic debilitating medical illnesses. Beneficial anabolic effects result on skin, muscle and bone in relation to the aging process with a concomitant decrease in body fat. Treatment of cancer patients by these peptides is also included, for example, prevention and/or reduction of cachexia in cancer patients. These therapeutic uses are accomplished by using a therapeutically effective amount of the compound. Such an amount is that needed to promote the release of serum growth hormone levels as discussed, infra.
The compounds of this invention may also be used to enhance blood GH levels in animals; enhance milk production in cows; enhance body growth in animals such as, e.g., humans, sheep, bovines, and swine, as well as fish, fowl, other vertebrates and crustaceans; and increase wool and/or fur production in mammals. The amount of body growth is dependent upon the sex and age of the animal species, quantity and identity of the growth hormone releasing compound being administered, route of administration, and the like.
Also, the compounds of this invention increase serum GH in humans; enhance body growth in short stature children; decrease body fat and improve protein metabolism in select children; improve protein metabolism of the skin, muscle, bone while decreasing body fat of the elderly, particularly when GH deficiency is present.
These compounds are also useful for improving serum lipid pattern in humans by decreasing in the serum the amount of serum cholesterol and low density lipoprotein, and increasing in the serum the amount of the high density lipoprotein.
The novel secretagogues of this invention can be synthesized according to the usual methods of solution and solid phase peptide chemistry, or by classical methods known in the art.
In accordance with another embodiment of the present invention, a method is provided for promoting release and/or elevation of growth hormone levels in the blood of an animal. This method of promoting the release and/or elevation of growth hormone levels can also be used to therapeutically treat the aforesaid diseases. Said methods comprise administering to an animal an effective dose of at least one of the above-described compounds. In one embodiment, this method is used in animals other than humans.
The compounds of this invention can be administered by oral, parenteral (intramuscular (i.m.), intraperitoneal (i.p.), intravenous (i.v.) or subcutaneous (s.c.) injection), nasal, vaginal, rectal or sublingual routes of administration as well as intrapulmonary inhalation can be formulated in dose forms appropriate for each route of administration. Parenteral administration is preferred.
Solid dose forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dose forms, the active compound is mixed with at least one inert carrier such as sucrose, lactose, or starch. Such dose forms can also comprise, as is normal practice, additional substances other than inert diluents, e.g., lubricating agents such as magnesium stearate. In the case of capsules, tablets and pills, the dose forms may also comprise buffering agents. Tablets and pills can additionally be prepared with enteric coatings.
Liquid dose forms for oral administration include emulsions, solutions, suspensions, syrups, the elixirs containing inert diluents commonly used in the art, such as water. Besides, such inert diluents, compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.
Preparations according to this invention for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, or emulsions. Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils, such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. Such dose forms may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. They may be sterilized by, for example, filtration through a bacteria-retaining filter, by incorporating sterilizing agents into the compositions, by irradiating the compositions, or by heating the compositions. They can also be manufactured in a medicum of sterile water, or some other sterile injectable medium immediately before use.
The amount of secretagogues or combination of compounds of the present invention administered will vary depending on numerous factors, e.g., the particular animal treated, its age and sex, the desired therapeutic affect, the route of administration and which polypeptide or combination of polypeptides are employed. In all instances, however, a dose effective (therapeutically effective amount) to promote release and elevation of growth hormone level in the blood of the recipient animal is used. Ordinarily, this dose level falls in the range of between about 0.1 xcexcg to 10 xcexcg of total compound per kg of body weight. The preferred amount can readily be determined empirically by the skilled artisan based upon the present disclosure.
For example, in humans when the mode of administration is i.v. the preferred dose level falls in the range of about 0.1 xcexcg to 10 xcexcg of total secretagogue per kg of body weight, more preferably, about 0.5 xcexcg to 5 xcexcg of total secretagogue per kg of body weight, still more preferably about 0.7 xcexcg about 3.0 xcexcg per kg of body weight. When combinations of growth hormone releasing compounds are used, lower amounts of the presently described peptide can be used. For example, combining the presently described secretagogues with, for example, a synergistic compound in Group I of U.S. Pat. No. 4,880,778 such as GHRH, or U.S. Pat. Nos. 5,663,146 or 5,486,505, a preferred range is about 0.1 xcexcg to about 5 xcexcg of the presently described compound per kg of body weight and about 0.5 xcexcg to about 15.04 xcexcg of synergistic compound (e.g. GHRH) and more preferably about 0.1 xcexcg to about 3 xcexcg of the present compound with about 1.0 xcexcg to about 3.0 xcexcg of the synergistic compound per kg of body weight.
When the mode of administration is oral, greater amounts are typically needed. For example, in humans for oral administration, the dose level is typically about 30 xcexcg to about 1200 xcexcg of compound per kg of body weight, more preferably about 70 xcexcg to about 600 xcexcg of compound per kg of body weight, still more preferably, about 200 xcexcg to about 600 xcexcg of total compound per kg of body weight. Cows and pigs require about the same dose level as humans, while rats typically require higher dose levels. The exact level can readily be determined empirically based upon the present disclosure.
In general, as aforesaid, the administration of combinations of growth hormone releasing peptides will allow for lower doses of the individual growth hormone releasing compounds to be employed relative to the dose levels required for individual growth hormone releasing compounds in order to obtain a similar response, due to the synergistic effect of the combination.
Also included within the scope of the present invention are compositions that comprise, as an active ingredient, the organic and inorganic addition salts of the above-described polypeptides and combinations thereof; optionally, in association with a carrier, diluent, slow release matrix, or coating.
The organic or inorganic addition salts of the growth hormone releasing compounds and combinations thereof contemplated to be within the scope of the present invention include salts of such organic moieties as acetate, trifluoroacetate, oxalate, valerate, oleate, laurate, benzoate, lactate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthalate, and the like; and such inorganic moieties as Group I (i.e., alkali metal salts), Group II (i.e. alkaline earth metal salts) ammonium and protamine salts, zinc, iron, and the like with counterions such as chloride, bromide, sulfate, phosphate and the like, as well as the organic moieties referred to above.
Pharmaceutically acceptable salts are preferred when administration to human subjects is contemplated. Such salts include the non-toxic alkali metal, alkaline earth metal and ammonium salts commonly used in the pharmaceutical industry including sodium, potassium, lithium, calcium, magnesium, barium, ammonium and protamine salts which are prepared by methods well known in the art. The term also includes non-toxic acid addition salts which are generally prepared by reacting the compounds of this invention with a suitable organic or inorganic acid. Representative salts include hydrochloride, hydrobromide, sulfate, bisulfate, acetate, oxalate, valerate, oleate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate and the like.